Research paper
Molecular docking and dynamic studies of a potential therapeutic target inhibiting glyoxalase system: Metabolic action of the 3, 3 '- [3- (5-chloro-2-hydroxyphenyl) -3-oxopropane-1, 1-diyl] - Bis-4-hydroxycoumarin leads overexpression of the intracellular level of methylglyoxal and induction of a pro-apoptotic phenomenon in a hepatocellular carcinoma model

https://doi.org/10.1016/j.cbi.2021.109511Get rights and content

Highlights

  • 1. The docking of OT-55 inside the active site of Glo-I indicates a potent binding.

  • OT-55 exhibits selective inhibitory effect on the glyoxalase system in vitro.

  • OT-55 induces pro-apoptotic process in DEN-initiated hepatocarcinoma.

  • In normal cells, it induces a vascular rearrangement.

  • 5. A preferential distribution of the encapsulated coumarin in liver was noted. .

Abstract

Methylglyoxal is a dicarbonyl compound recruited as a potential cytotoxic marker, initially presents in cells and considered as a metabolite of the glycolytic pathway. Our aim is to demonstrate the inhibitory effect of 3, 3'-[3-(5-chloro-2-hydroxyphenyl)-3-oxopropane-1, 1-diyl] Bis (4-hydroxycoumarin) on the glyoxalase system, and indirectly its anticancer activity. The docking of OT-55 was conducted by using Flexible docking protocol, ChiFlex and libdock tools inside the active site of Glo-I indicated that both hydrogen bonding and hydrophobic interactions contributed significantly in establishing potent binding with the active site which is selected as a strong inhibitor with high scoring values and maximum Gibbs free energy. Coumarin-liposome formulation was characterized and evaluated in vivo against chemically induced hepatocarcinoma in Wistar rats. After Diethylnitrosamine (DEN) induction, microscopic assessment was realized; precancerous lesions were developed showing an increase of both tumor-associated lymphocyte and multiple tumor acini supported by the blood investigation. Our finding also suggested a preferential uptake of liposomes respectively in liver, kidney, lung, brain and spleen in the DEN-treated animals. OT-55 has also been shown to inhibit the activity of Glo-I in vitro as well as in DEN-treated rats. An abnormal high level of MGO of up to 50% was recorded followed by a reduction in glucose consumption and lactate dehydrogenase production validated in the positive control. MGO generates apoptosis as depicted by focal hepatic lesions. Also, no deleterious effects in the control group were observed after testing our coumarin but rather a vascular reorganization leading to nodular regenerative hyperplasia. Involved in the detoxification process, liver GSH is restored in intoxicated rats, while no changes are seen between controls. At the endothelial cell, OT-55 appears to modulate the release of NO only in the DEN-treated group. OT-55 would behave both as an anticancer agent but also as an angiogenic factor regarding results obtained.

Introduction

The World Health Organization reports reveal that cancer is a real public health problem because around 56 million deaths were recorded in 2000, of which 12% were associated with malignant tumors. According to the same source, 15 million new cases will appear each year until 2020 (http://www.who.int/mediacentre/news/releases/2003/pr27/fr/press release). This alarming increase of new patients with cancer entails health sectors to take proactive measures to cease this upsurge. The numbers associated to this pathology situate liver cancer at the third position (8.8%) after lung (17.8%) and stomach (10.4%). Actually, the magnetic resonance imaging (MRI) is one of the most extensively used methods in detection of hepatocellular diseases and tumor metastasis from liver and other organs [1]. Unfortunately, this type of tool is not always available which hinders the diagnosis. In addition, the epigenetic nature of cancer makes diagnosis difficult because of the molecular mechanism modulating the tumor expression genes. The use of conventional therapies (chemotherapy, radiotherapy etc.) has reached their limits in terms of efficiency. New nanoparticle systems in tumor targeting have been proposed. These vehicles have been used to reduce toxicity and modify the pharmacokinetics of anticancer drugs carriers [2]. In fact, it is estimated that more than 90% of cancer drugs exhibit poor bioavailability [3]. One of the classic examples is liposomal doxorubicin used to treat ovarian cancer which has undergone clinical approval [4] and liposomal Cytarabine used to treat lymphomatous meningitis [5]. In this article, Coumarin-liposomal encapsulation was used mainly to overcome the inevitable problem of both solubility and toxicity. Our work shows that natural coumarins, although abundant, have not finished surprising us. Coumarins and their derivatives are getting importance because of their pharmacological activities such as anti-bacterial, anti-thrombotic and vasodilatory, antimutagenic, lipoxygenase and cyclooxygenase inhibition, scavenging of reactive oxygen species, and anti-tumorigenic [6]. In fact, the various substituted coumarins that have been synthesized have had a double impact by strengthening the structure while improving its therapeutic effects (anticoagulant, anti-inflammatory, anti-HIV, antioxidant, anti-allergic, anti-cancer, anti-proliferative and antiviral activities) as described by Pandey [7]. In this context, we have targeting the cytotoxicity of 3, 3 '- [3- (5-chloro-2-hydroxyphenyl) -3- oxopropane-1, 1-diyl] - bis-4-hydroxycoumarin (OT-55), a new synthetic compound by blocking the conversion of methylglyoxal (MGO) to d-Lactate via the intermediate S-d-lactoylglutathione [8].

Using this anti-cancer strategy involving methylglyoxal seems to be nimble as it is a physiological metabolic side-product that is principally produced within glycolysis reaction [9]. In liver, the site of the carbohydrate metabolism, the overexpression of glyoxalase system (Glo-1 and Glo-2) provides the primary defense against dicarbonyl stress (MGO) at the origin of the glycation phenomenon (AGE). The abnormal accumulation of dicarbonyl metabolites leads to increase in protein and DNA mutations which contribute to cell and tissue dysfunction in ageing and disease [10].Therefore; it is responsible for methylglyoxal detoxification [11]. Discovery of the critical role of MGO has opened the doors to many speculations on the best way to get around this problem since it is demonstrated its selectivity toward cancer cells [12] without affecting the normal cells. Thereby, it is clearly established the indirect implication of MGO levels as a precursor of advanced glycation end product in the experimental diabetic nephropathy [13]. Furthermore, it was reported that methylglyoxal Bis-guanylhydrazone (MGBG) used as an antineoplastic agent, has a useful treatment for intracerebral tumors [14]. In addition, an increasing number of studies have identified the tumorigenic potential of the MGO in breast cancer cells by activating the MAPK pathway and down-regulating the Bcl2 and MMP-9 [15].

Taken together; the molecular docking and in vitro monitoring reaction of OT-55 against GLO I, allowed us to show the indirect effect of dicarbonyl stress (MGO) on Wistar rats with hepatocarcinoma caused by diethylnitrosamine (DEN). This conventional but mandatory approach seemed necessary to demonstrate the cytotoxic properties of OT-55 on cancer cells. For better pharmacokinetic characterization and tissue distribution of the prepared coumarin-liposome, we evaluated several parameters such as methylglyoxal, OT-55, GSH and NO in plasma and tissues before and after treatment. Finally, a complete liver assessment has also been established.

Section snippets

Reagent and standards

Recombinant Human Glyoxalase I Protein, CF (NP _006699) was purchased from R & D Systems Bio-Tech, Minneapolis MN, USA. l-Glutathione-Reduced (γ-L-glutamyl-L-cysteinyl-glycine, GSH), Methylglyoxal solution (acetylformaldehyde, pyruvaldehyde, pyruvic aldehyde), N-Nitrosodiethylamine, carbon tetrachloride, analytical standard of 2-Methylquinoxaline, O-Phenylenediamine were delivered by Sigma-Aldrich-Portugal. Stainless Steel Needle for rats: FTSS-13ga (1.3 × 1.7 mm) was purchased from Instech,

Docking of OT-55 against Glo-I

The docking of OT-55 has resulted in producing 189 poses and pattern of binding with CDOCKER scores ranging from 39.6 to 106.3 and a Libdock score from 39.5 to 117.1. The best scoring compound was investigated and it was found fitting very well in the active site and forming a series of essential bonding that explain its high scoring values.

Fig. 1 shows 3D and 2D representation of OT-55 inside the active site. Interestingly, one of the chromen rings is fitting well next the zinc atom in which

Discussion

In this paper the in vitro and in vivo therapeutic effect of the OT-55 by using conventional technique i.e., drug-loaded liposomes was investigated against DEN-initiated hepatocarcinogesesis via the glycolytic intermediates: the methylglyoxal. We have noticed during the various solvation tests that the coumarin (OT-55) was lipohilic and intercalated between the lipid bilayer as described previously by Roy[24] and confirmed by ATR-FTIR spectrum (supplementary data) and NMR, a part nevertheless

5. Conclusion

Finally, it is expected that starving cancer cells by cycling the glycolytic pathway seems the only alternative to thwart the undesirable effects of unintended multiplication of cancer cells.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Conflicts of interest

The authors declare that they have no conflict of interest.

Ethics approval

All animal experiments were carried out in accordance with the U.K. Animals (Scientific Procedures) Act, 1986 and associated guidelines, EU Directive 2010/63/EU for animal experiments: http://ec.europa.eu/environment/chemicals/lab animals/legislation en.htm.

Availability of data and material

Supplementary Data information is available for this paper.

Author contributions

N.Taïbi contributed to the conception and design of the study (tumor induction and treatment, liposome synthesis and characterization, optimization of analysis methods and drafting the version of paper); A. Silva supervised the project and participated to English correction; N. Bekari contributed to hepatocarcinoma induction and treatment; O.Talhi and L. Boukenna participated to OT-55 synthesis and their NMR characterization; R. Ameraoui, M. Hadjadj, M. Abou-Mustapha, Z. M. Boutaiba, F. Dergal,

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgements

We would like to thank the technical assistance of electron microscopy laboratory from Faculty of Biology of the University of Science and Technology Houari Boumediene (USTHB).

References (69)

  • G. Renier et al.

    Effect of gliclazide on monocyte-endothelium interactions in diabetes

    J. Diabetes Complicat.

    (2000)
  • P.P. Hsu et al.

    Cancer cell metabolism: warburg and beyond

    Cell

    (2008)
  • P.J. Thornalley

    The glyoxalase system in health and disease

    Mol. Aspect. Med.

    (1993)
  • R. Dringen

    Metabolism and functions of glutathione in brain

    Prog. Neurobiol.

    (2000)
  • N. Shangari et al.

    The cytotoxic mechanism of glyoxal involves oxidative stress

    Biochem. Pharmacol.

    (2004)
  • C.Y. Tu et al.

    Methylglyoxal induces DNA crosslinks in ECV304 cells via a reactive oxygen species-independent protein carbonylation pathway

    Toxicol. Vitro

    (2013)
  • L.H. Lv et al.

    Anticancer drugs cause release of exosomes with heat shock proteins from human hepatocellular carcinoma cells that elicit effective natural killer cell antitumor responses in vitro

    J. Biol. Chem.

    (2012)
  • A.M. Slavotinek et al.

    Unfolding the role of chaperones and chaperonins in human disease

    Trends Genet.

    (2001)
  • T.A. Cowger et al.

    Casein-coated Fe5C2 nanoparticles with superior r2 relaxivity for liver-specific magnetic resonance imaging

    THERANOSTICS

    (2015)
  • I. Takuya

    Challenges of drug delivery systems that contribute to cancer chemotherapy

    Biol. Pharm. Bull.

    (2013)
  • S. Stewart et al.

    Randomized comparative trial of pegylated liposomal doxorubicin versus bleomycin and vincristine in theTreatment of AIDS-related kaposi's sarcoma

    J CLIN ONC

    (1998)
  • M.J. Glantz et al.

    Randomized trial of a slow-release versus a StandardFormulation of cytarabine for the intrathecal treatmentof lymphomatous meningitis

    J CLIN ONC

    (1991)
  • M. Ismail et al.

    Chemical constituents and antioxidant activity of Geranium wallichianum

    Record Nat. Prod.

    (2009)
  • H. Pandey et al.

    Liposome and their applications in cancer therapy

    Braz. Arch. Biol. Technol.

    (2016)
  • Q.A. Al-Balas et al.

    Novel glyoxalase-I inhibitors possessing a "zinc-binding feature" as potential anticancer agents

    Drug Des. Dev. Ther.

    (2016)
  • S.A. Philips et al.

    The formation of methylglyoxal from triose phosphates

    Eur. J. Biochem.

    (1993)
  • N. Rabbani et al.

    Dicarbonyls and glyoxalase in disease mechanisms and clinical therapeutics

    GLYCO J

    (2016)
  • K. Seo et al.

    Methylglyoxal induces mitochondrial dysfunction and cell death in liver

    TOXICOL RES

    (2014)
  • U.K. Biswas et al.

    Elevation of serum methylglyoxal may be used as a screening marker in oral premalignant lesions

    Biomed. Res.

    (2011)
  • N. Rabbani et al.

    The critical role of methylglyoxal and glyoxalase 1 in diabetic nephropathy

    Diabetes

    (2014)
  • M.G. Rosenblum et al.

    Penetration of methylglyoxal bis(guanylhydrazone) into intracerebral tumors in humans

    Can. Res.

    (1981)
  • Y. Guo et al.

    Effects of methylglyoxal and glyoxalase I inhibition on breast cancer cells proliferation, invasion, and apoptosis through modulation of MAPKs, MMP9, and Bcl-2

    CAN BIOL AND THER

    (2016)
  • E.C.M. Cabral et al.

    Reparation and characterization of liposomes entrapping allergenic proteins

    Braz. J. Chem. Eng.

    (2004)
  • Z. Hassannejad et al.

    Fabrication and characterization of magnetoplasmonic liposome carriers

    NANOSCIENCE & TECHNOLOGY

    (2014)
  • Cited by (0)

    View full text